論文 - 詳細
| RRC ID | 77371 |
|---|---|
| 著者 | Takakura H, Kojima R, Urano Y, Terai T, Hanaoka K, Nagano T. |
| タイトル | Aminoluciferins as functional bioluminogenic substrates of firefly luciferase. |
| ジャーナル | Chem Asian J |
| Abstract |
Firefly luciferase is widely used as a reporter gene in assays to study gene expression, gene delivery, and so on because of its extremely high signal-to-noise ratio. The availability of a range of bioluminogenic substrates would greatly extend the applicability of the luciferin-luciferase system. Herein, we describe a design concept for functional bioluminogenic substrates based on the aminoluciferin (AL) scaffold, together with a convenient, high-yield method for synthesizing N-alkylated ALs. We confirmed the usefulness of ALs as bioluminogenic substrates by synthesizing three probes. The first was a conjugate of AL with glutamate, Glu-AL. When Glu-AL, the first membrane-impermeable bioluminogenic substrate of luciferases, was applied to cells transfected with luciferase, luminescence was not observed; that is, by using Glu-AL, we can distinguish between intracellular and extracellular events. The second was Cy5-AL, which consisted of Cy5, a near-infrared (NIR) cyanine fluorescent dye, and AL, and emitted NIR light. When Cy5-AL reacted with luciferase, luminescence derived from Cy5 was observed as a result of bioluminescence resonance energy transfer (BRET) from AL to Cy5. The NIR emission wavelength would allow a signal to be observed from deeper tissues in bioluminescence in vivo imaging. The third was biotin-DEVD-AL (DEVD = the amino acid sequence Asp-Glu-Val-Asp), which employed a caspase-3 substrate peptide as a switch to control the accessibility of the substrate to luciferase, and could detect the activity of caspase-3 in a time-dependent manner. This generalized design strategy should be applicable to other proteases. Our results indicate that the AL scaffold is appropriate for a range of functional luminophores and represents a useful alternative substrate to luciferin. |
| 巻・号 | 6(7) |
| ページ | 1800-10 |
| 公開日 | 2011-7-4 |
| DOI | 10.1002/asia.201000873 |
| PMID | 21416616 |
| MeSH | Biosensing Techniques Biotin / chemistry Caspase 3 / metabolism Firefly Luciferin / chemistry* Fluorescence Resonance Energy Transfer Fluorescent Dyes / chemistry* Luciferases, Firefly / chemistry* Luciferases, Firefly / metabolism Spectroscopy, Near-Infrared Substrate Specificity |
| IF | 4.056 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 9 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | COS-7(RCB0539) |